低生物量微生物组研究中污染预防与报告指南

Guidelines for preventing and reporting contamination in low-biomass microbiome studies.

作者信息

Fierer Noah, Leung Pok Man, Lappan Rachael, Eisenhofer Raphael, Ricci Francesco, Holland Sophie I, Dragone Nicholas, Blackall Linda L, Dong Xiyang, Dorador Cristina, Ferrari Belinda C, Goordial Jacqueline, Holmes Susan P, Inagaki Fumio, Korem Tal, Li Simone S, Makhalanyane Thulani P, Metcalf Jessica L, Nagarajan Niranjan, Orsi William D, Shanahan Erin R, Walker Alan W, Weyrich Laura S, Gilbert Jack A, Willis Amy D, Callahan Benjamin J, Shade Ashley, Parkhill Julian, Banfield Jillian F, Greening Chris

机构信息

Department of Ecology and Evolutionary Biology, Cooperative Institute for Research in Environmental Sciences, University of Colorado Boulder, Boulder, CO, USA.

Department of Microbiology, Biomedicine Discovery Institute, Monash University, Clayton, Victoria, Australia.

出版信息

Nat Microbiol. 2025 Jun 20. doi: 10.1038/s41564-025-02035-2.

Abstract

Numerous important environments harbour low levels of microbial biomass, including certain human tissues, the atmosphere, plant seeds, treated drinking water, hyper-arid soils and the deep subsurface, with some environments lacking resident microbes altogether. These low microbial biomass environments pose unique challenges for standard DNA-based sequencing approaches, as the inevitability of contamination from external sources becomes a critical concern when working near the limits of detection. Likewise, lower-biomass samples can be disproportionately impacted by cross-contamination and practices suitable for handling higher-biomass samples may produce misleading results when applied to lower microbial biomass samples. This Consensus Statement outlines strategies to reduce contamination and cross-contamination, focusing on marker gene and metagenomic analyses. We also provide minimal standards for reporting contamination information and removal workflows. Considerations must be made at every study stage, from sample collection and handling through data analysis and reporting to reduce and identify contaminants. We urge researchers to adopt these recommendations when designing, implementing and reporting microbiome studies, especially those conducted in low-biomass systems.

摘要

许多重要环境中微生物生物量水平较低,包括某些人体组织、大气、植物种子、处理过的饮用水、超干旱土壤和深层地下环境,有些环境甚至完全没有常驻微生物。这些低微生物生物量环境给基于DNA的标准测序方法带来了独特挑战,因为在接近检测极限时,不可避免的外源污染成为关键问题。同样,低生物量样本可能受到交叉污染的影响更大,适用于处理高生物量样本的操作应用于低微生物生物量样本时可能会产生误导性结果。本共识声明概述了减少污染和交叉污染的策略,重点关注标记基因和宏基因组分析。我们还提供了报告污染信息和去除工作流程的最低标准。在从样本采集、处理到数据分析和报告的每个研究阶段都必须加以考虑,以减少和识别污染物。我们敦促研究人员在设计、实施和报告微生物组研究时采用这些建议,特别是在低生物量系统中进行的研究。

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